Concentration Quantification of the Low-Complexity Domain of Fused in Sarcoma inside a Single Droplet and Effects of Solution Parameters
Concentration Quantification of the Low-Complexity Domain of Fused in Sarcoma inside a Single Droplet and Effects of Solution Parameters
复制标题
单液滴内肉瘤融合低复杂域的浓度定量及溶液参数的影响
DOI：
10.1021/acs.jpclett.2c00962
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Nakabayashi Takakazu
中科院分区：
文献类型：
--
作者：
Yokosawa Kohei;Kajimoto Shinji;Shibata Daiki;Kuroi Kunisato;Konno Tomohiro;Nakabayashi Takakazu
液-液相分离（LLPS）是生物学中的一种重要现象，因此需要开发定量分析LLPS产生的蛋白液滴的方法。本研究使用拉曼成像和水的拉曼谱带作为强度标准，量化了无标记和单液滴条件下液滴中蛋白质浓度的变化。观察到蛋白质浓度随pH和盐浓度变化的微小变化，并且表明液滴中的浓度随着条件变得不利于液滴形成而降低。还检查了暴露于1，6-己二醇的效果，发现该添加剂降低了液滴中的蛋白质浓度。可以提出一种模型，其中加入1，6-己二醇降低了液滴中的蛋白质浓度，并且当浓度福尔斯到某一阈值以下时液滴消失。
Liquid–liquid phase separation (LLPS) is an important phenomenon in biology, and it is desirable to develop quantitative methods to analyze protein droplets generated by LLPS. This study quantified the change in protein concentration in a droplet in label-free and single-droplet conditions using Raman imaging and the Raman band of water as an intensity standard. Small changes in the protein concentration with variations in pH and salt concentration were observed, and it was shown that the concentration in the droplet decreases as the conditions become less favorable for droplet formation. The effect of exposure to 1,6-hexanediol was also examined, and this additive was found to decrease the protein concentration in the droplet. A model can be proposed in which the addition of 1,6-hexanediol reduces the protein concentration in the droplet, and the droplet disappears when the concentration falls below a certain threshold value.